Fig. 5

In vitro functional validation of JQ1 attenuation of PDGF and TGFB stimulated contraction in RBMC. A Representative images of RBMC stimulated with PBS (Veh), PDGF, TGFB with DMSO or JQ1 for 16 h and stained with Vimentin, Phalloidin and DAPI. B Phalloidin intensity and cell shape was quantified in ImageJ. C RBMC were plated on 1.2 mg/mL collagen gels in a 12 well plate in 1 mL of serum free media and incubated for 16 h with PBS (Veh), PDGF, TGFB with DMSO or JQ1. Select wells were stimulated for 30 min with a ROCK inhibitor as a negative control for contraction. Collagen gels were separated from the walls of the wells and Imaged after 1.5 h to capture spontaneous (unstimulated contraction, followed by an additional 1.5 h in 5% FBS to facilitate contraction. D Contraction was quantified using ImageJ and measured as a percent of the changed area of the gel from the baseline. Data are representative of 3 independent trials. 1NC_FBS refers to negative control for FBS in which cells never receive FBS from platting to harvesting. 2NC_iROCK refers to the negative control for contraction in which cells are stimulated with a ROCK inhibitor for 30 min prior to observing contraction. *, p < 0.05, ** p < 0.01, ***, p < 0.001, ****, p < 0.0001 compared to control. #, p < 0.05, ## p < 0.01, ###, p < 0.001, ###, p < 0.0001 compared to PDGF. $, p < 0.05, $$ p < 0.01, $$$, p < 0.001, $$$$, p < 0.0001 compared to TGFB. Statistical significance was calculated with student t-test